Methods and systems of diagnosing brain injury

Abstract

Disclosed herein are methods and systems of determining whether a subject's levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated in a sample collected from the subject. The methods comprise determining whether the levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated in the sample, and communicating the determination on or from an instrument. The methods may be used to aid in the diagnosis and evaluation of a subject (e.g., a human subject) that has sustained or may have sustained an injury to the head, such as to determine whether the subject is suffering from a mild, moderate, severe, or moderate to severe traumatic brain injury (TBI).

What is claimed is: 
     
         1 . A method comprising the steps of:
 a. performing at least one assay for ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) and at least one assay for glial fibrillary acidic protein (GFAP) in at least one sample obtained from a human subject, wherein the sample is obtained from the subject within about 12 to within about 48 hours after an actual or suspected injury to the head;   b. determining that:
 1. the subject's levels of GFAP and UCH-L1 are elevated when (i) the level of GFAP in the sample is equal to or above about 30 pg/mL and level of UCH-L1 in the sample is below about 360 pg/mL, cannot be determined or is not reported; (ii) the level of GFAP in the sample is equal to or above about 30 pg/mL and level of UCH-L1 in the sample is equal to or above about 360 pg/mL; or (iii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is equal to or above about 360 pg/mL; 
 2. the subject's levels of GFAP and UCH-L1 are not elevated when the level of GFAP in the sample is below about 30 pg/mL and the level of UCH-L1 in the sample is below about 360 pg/mL; or 
 3. the assays for UCH-L1 and GFAP should be repeated when (i) the level of GFAP in the sample is below about 30 pg/mL and the level of UCH-L1 in the sample cannot be determined or is not reported; (ii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is below about 360 pg/mL; or (iii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample cannot be determined or is not reported, and 
   c. communicating the determination from step b (1)-(3) on or from at least one instrument, wherein the instrument is a point-of-care device.   
     
     
         2 . The method of  claim 1 , wherein the method further comprises:
 a. performing a head computed tomography (CT) scan, magnetic resonance imaging (MRI) procedure, or both a CT scan or a MRI procedure on the subject when the subject's levels of GFAP and UCH-L1 are elevated, or   b. not performing a head CT scan or an MRI procedure on the subject when the subject's levels of GFAP and UCH-L1 are not elevated.   
     
     
         3 . The method of  claim 1 , further comprising diagnosing the subject as having a traumatic brain injury (TBI) when the level of GFAP is equal to or above about 30 pg/mL and the level of UCH-L1 is equal to or above about 360 pg/mL, regardless of whether a head CT scan is negative for a TBI or whether any head CT scan is performed. 
     
     
         4 . The method of  claim 1 , wherein the method further comprises:
 (a) treating the subject for a mild, moderate, moderate to severe, or severe TBI when the subject's levels of GFAP and UCH-L1 are elevated;   (b) monitoring the subject when the subject's levels of GFAP and UCH-L1 are elevated; or   (c) a combination of (a) and (b).   
     
     
         5 . (canceled) 
     
     
         6 . The method of  claim 1 , wherein the sample is taken within about 13 hours to within about 48 hours, within about 14 hours to within about 48 hours, within about 15 hours to within about 48 hours, within about 16 hours to within about 48 hours, within about 17 hours to within about 48 hours, within about 18 hours to within about 48 hours, within about 19 hours to within about 48 hours, within about 20 hours to within about 48 hours, within about 21 hours to within about 48 hours, within about 22 hours to within about 48 hours, within about 23 hours to within about 48 hours, within about 24 hours to within about 48 hours, 25 hours to within about 48 hours, within about 26 hours to within about 48 hours, within about 27 hours to within about 48 hours, within about 29 hours to within about 48 hours, within about 30 hours to within about 48 hours, within about 31 hours to within about 48 hours, within about 32 hours to within about 48 hours, within about 33 hours to within about 48 hours, within about 34 hours to within about 48 hours, within about 35 hours to within about 48 hours, within about 36 hours to within about 48 hours, within about 37 hours to within about 48 hours, within about 38 hours to within about 48 hours, within about 39 hours to within about 48 hours, within about 40 hours to within about 48 hours, or within about 12 hour to within about 48 hours after the actual or suspected injury to the head. 
     
     
         7 . (canceled) 
     
     
         8 . The method of  claim 1 , wherein the sample is obtained:
 (a) after the subject sustained an injury to the head caused by physical shaking, blunt impact by an external mechanical or other force that results in a closed or open head trauma, one or more falls, explosions or blasts or other types of blunt force trauma;   (b) after the subject has ingested or been exposed to a chemical, toxin or combination of a chemical and toxin; or   (c) from a subject that suffers from an autoimmune disease, a metabolic disorder, a brain tumor, hypoxia, a viral infection, a fungal infection, a bacterial infection, meningitis, hydrocephalus, or any combinations thereof.   
     
     
         9 . (canceled) 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . The method of  claim 1 , wherein the assay is (a) an immunoassay or a clinical chemistry assay; or (b) a single molecule detection assay or a point-of-care assay. 
     
     
         13 . (canceled) 
     
     
         14 . The method of  claim 1 , wherein the amount of the at least one sample is: (a) about 10 μL to about 30 μL; or (b) about 20 μL. 
     
     
         15 . (canceled) 
     
     
         16 . The method of  claim 1 , wherein the at least one assay for UCH-L1, at least one assay for GFAP, or at least one assay for UCH-L1 and at least one assay for GFAP is performed in: (a) about 10 to about 20 minutes; or (b) about 15 minutes. 
     
     
         17 . (canceled) 
     
     
         18 . (canceled) 
     
     
         19 . The method of  claim 1 , wherein the sample is selected from the group consisting of a whole blood sample, a serum sample, and a plasma sample. 
     
     
         20 . A system comprising:
 an assay for ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) and an assay for glial fibrillary acidic protein (GFAP); and   a point-of-care device for performing the assay for UCH-L1 and the assay for GFAP, wherein   the device determines an amount of UCH-L1 and GFAP in a sample obtained from a subject, wherein the sample is obtained from the subject within about 12 to within about 48 hours after an actual or suspected injury to the head, and   the amount of UCH-L1 and GFAP determined in the sample are communicated on or from the device as:
 a. elevated when (i) the level of GFAP in the sample is equal to or above about 30 pg/mL and level of UCH-L1 in the sample is below about 360 pg/mL, cannot be determined or is not reported; (ii) the level of GFAP in the sample is equal to or above about 30 pg/mL and level of UCH-L1 in the sample is equal to or above about 360 pg/mL; or (iii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is equal to or above about 360 pg/mL; 
 b. not elevated when the level of GFAP in the sample is below about 30 pg/mL and level of UCH-L1 in the sample is below about 360 pg/mL; or 
 c. requiring the assays for UCH-L1 and GFAP to be repeated when (i) the level of GFAP in the sample is below about 30 pg/mL and the level of UCH-L1 in the sample cannot be determined or is not reported; (ii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is below about 360 pg/mL; or (iii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample cannot be determined or is not reported. 
   
     
     
         21 . The system of  claim 20 , wherein the sample is taken within about 12 hours to within about 48 hours, within about 13 hours to within about 48 hours, within about 14 hours to within about 48 hours, within about 15 hours to within about 48 hours, within about 16 hours to within about 48 hours, within about 17 hours to within about 48 hours, within about 18 hours to within about 48 hours, within about 19 hours to within about 48 hours, within about 20 hours to within about 48 hours, within about 21 hours to within about 48 hours, within about 22 hours to within about 48 hours, within about 23 hours to within about 48 hours, within about 24 hours to within about 48 hours, 25 hours to within about 48 hours, within about 26 hours to within about 48 hours, within about 27 hours to within about 48 hours, within about 29 hours to within about 48 hours, within about 30 hours to within about 48 hours, within about 31 hours to within about 48 hours, within about 32 hours to within about 48 hours, within about 33 hours to within about 48 hours, within about 34 hours to within about 48 hours, within about 35 hours to within about 48 hours, within about 36 hours to within about 48 hours, within about 37 hours to within about 48 hours, within about 38 hours to within about 48 hours, within about 39 hours to within about 48 hours, within about 40 hours to within about 48 hours, or within about 12 hour to within about 48 hours after an actual or suspected injury to the head. 
     
     
         22 . (canceled) 
     
     
         23 . The system of  claim 20 , wherein the sample is obtained:
 (a) after the subject sustained an injury to the head caused by physical shaking, blunt impact by an external mechanical or other force that results in a closed or open head trauma, one or more falls, explosions or blasts or other types of blunt force trauma;   (b) after the subject has ingested or been exposed to a chemical, toxin or combination of a chemical and toxin; or   (c) from a subject that suffers from an autoimmune disease, a metabolic disorder, a brain tumor, hypoxia, a viral infection, a fungal infection, a bacterial infection, meningitis, hydrocephalus, or any combinations thereof.   
     
     
         24 . (canceled) 
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . The system of  claim 20 , wherein the assay is (a) an immunoassay or a clinical chemistry assay; or (b) a single molecule detection assay or a point-of-care assay. 
     
     
         28 . (canceled) 
     
     
         29 . The system of  claim 20 , wherein the amount of the at least one sample is: (a) about 10 μL to about 30 μL; or (b) about 20 μL. 
     
     
         30 . (canceled) 
     
     
         31 . The system  claim 20 , wherein the at least one assay for UCH-L1, at least one assay for GFAP, or at least one assay for UCH-L1 and at least one assay for GFAP is performed in (a) about 10 to about 20 minutes; or (b) about 15 minutes. 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . The system of  claim 20 , wherein the sample is selected from the group consisting of a whole blood sample, a serum sample, and a plasma sample. 
     
     
         35 . A method comprising the steps of:
 a. performing at least one assay for ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), at least one assay for glial fibrillary acidic protein (GFAP), or at least one assay for UCH-L1 and at least one assay for GFAP, in at least one sample obtained from a human subject, wherein the sample is obtained from the subject within about 12 to within about 48 hours after an actual or suspected injury to the head;   b. determining that:
 1. the subject's levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated when (i) the level of GFAP alone in the sample is equal to or above about 30 pg/mL; (ii) the level of GFAP in the sample is equal to or above about 30 pg/mL and level of UCH-L1 in the sample is below about 360 pg/mL, cannot be determined or is not reported; (iii) the level of GFAP in the sample is equal to or above about 30 pg/mL and level of UCH-L1 in the sample is equal to or above about 360 pg/mL; (iv) the level of UCH-L1 alone in the sample is equal to or above 360 pg/mL; or (v) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is equal to or above about 360 pg/mL; 
 2. the subject's levels of (i) GFAP are not elevated when GFAP alone in the sample is below about 30 pg/mL; (ii) UCH-L1 are not elevated when UCH-L1 alone in the sample is below about 360 pg/mL; or (iii) GFAP and UCH-L1 are not elevated when the level of GFAP in the sample is below about 30 pg/mL and level of UCH-L1 in the sample is below about 360 pg/mL; or 
 3. the assays for UCH-L1 and GFAP should be repeated when (i) the level of UCH-L1 alone in the sample cannot be determined or is not reported; (ii) the level of GFAP in the sample is below about 30 pg/mL and the level of UCH-L1 in the sample cannot be determined or is not reported; (iii) the level of GFAP alone in the sample cannot be determined or is not reported; (iv) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is below about 360 pg/mL; or (v) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample cannot be determined or is not reported, and 
   c. communicating the determination from step b (1)-(3) on or from at least one instrument, wherein the instrument is a point-of-care device.   
     
     
         36 . The method of  claim 35 , wherein the method further comprises:
 a. performing a head computed tomography (CT) scan, magnetic resonance imaging (MRI) procedure, or both a CT scan or a MRI procedure on the subject when the subject's levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated, or   b. not performing a head CT scan or an MRI procedure on the subject when the subject's levels of GFAP, UCH-L1, or GFAP and UCH-L1 are not elevated.   
     
     
         37 . The method of  claim 35 , further comprising diagnosing the subject as having a traumatic brain injury (TBI) when the level of GFAP is equal to or above about 30 pg/mL, the level of UCH-L1 is equal to or above about 360 pg/mL, or level of GFAP is equal to or above about 30 pg/mL and the level of UCH-L1 is equal to or above about 360 pg/mL, regardless of whether a head CT scan is negative for a TBI or whether any head CT scan is performed. 
     
     
         38 . The method of  claim 35 , wherein the method further comprises:
 (a) treating the subject for a mild, moderate, moderate to severe, or severe TBI when the subject's levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated;   (b) monitoring the subject when the subject's levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated; or   (c) a combination of (a) or (b).   
     
     
         39 . (canceled) 
     
     
         40 . The method of  claim 35 , wherein the sample is taken wherein the sample is taken within about 13 hours to within about 48 hours, within about 14 hours to within about 48 hours, within about 15 hours to within about 48 hours, within about 16 hours to within about 48 hours, within about 17 hours to within about 48 hours, within about 18 hours to within about 48 hours, within about 19 hours to within about 48 hours, within about 20 hours to within about 48 hours, within about 21 hours to within about 48 hours, within about 22 hours to within about 48 hours, within about 23 hours to within about 48 hours, within about 24 hours to within about 48 hours, 25 hours to within about 48 hours, within about 26 hours to within about 48 hours, within about 27 hours to within about 48 hours, within about 29 hours to within about 48 hours, within about 30 hours to within about 48 hours, within about 31 hours to within about 48 hours, within about 32 hours to within about 48 hours, within about 33 hours to within about 48 hours, within about 34 hours to within about 48 hours, within about 35 hours to within about 48 hours, within about 36 hours to within about 48 hours, within about 37 hours to within about 48 hours, within about 38 hours to within about 48 hours, within about 39 hours to within about 48 hours, within about 40 hours to within about 48 hours, or within about 12 hour to within about 48 hours after the actual or suspected injury to the head. 
     
     
         41 . (canceled) 
     
     
         42 . The method of  claim 35 , wherein the sample is obtained:
 (a) after the subject sustained an injury to the head caused by physical shaking, blunt impact by an external mechanical or other force that results in a closed or open head trauma, one or more falls, explosions or blasts or other types of blunt force trauma;   (b) after the subject has ingested or been exposed to a chemical, toxin or combination of a chemical and toxin; or   (c) from a subject that suffers from an autoimmune disease, a metabolic disorder, a brain tumor, hypoxia, a viral infection, a fungal infection, a bacterial infection, meningitis, hydrocephalus, or any combinations thereof.   
     
     
         43 . (canceled) 
     
     
         44 . (canceled) 
     
     
         45 . (canceled) 
     
     
         46 . The method of  claim 35 , wherein the assay is (a) an immunoassay or a clinical chemistry assay; or (b) a single molecule detection assay or a point-of-care assay. 
     
     
         47 . (canceled) 
     
     
         48 . The method of  claim 35 , wherein the amount of the at least one sample is about 10 μL to about 30 μL; or (b) about 20 μL. 
     
     
         49 . (canceled) 
     
     
         50 . The method of  claim 35 , wherein the at least one assay for UCH-L1, at least one assay for GFAP, or at least one assay for UCH-L1 and at least one assay for GFAP is performed in: (a) about 10 to about 20 minutes; or (b) about 15 minutes. 
     
     
         51 . (canceled) 
     
     
         52 . (canceled) 
     
     
         53 . The method of  claim 35 , wherein the sample is selected from the group consisting of a whole blood sample, a serum sample, and a plasma sample. 
     
     
         54 . A system comprising:
 an assay for ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), an assay for glial fibrillary acidic protein (GFAP), or an assay for UCH-L1 and an assay for GFAP; and   a point-of-care device for performing the assay for UCH-L1, the assay for GFAP, or the assay for UCH-L1 and the assay for GFAP, wherein   the device determines an amount of UCH-L1, GFAP, or UCH-L1 and GFAP in a sample obtained from a subject, wherein the sample is taken within about 12 hours to within about 48 hours; and   the amount of UCH-L1, GFAP, or UCH-L1 and GFAP determined in the sample are communicated on or from the device as:
 a. elevated when (i) the level of GFAP alone in the sample is equal to or above about 30 pg/mL; (ii) the level of GFAP in the sample is equal to or above about 30 pg/mL and level of UCH-L1 in the sample is below about 360 pg/mL, cannot be determined or is not reported; (iii) the level of GFAP in the sample is equal to or above about 30 pg/mL and level of UCH-L1 in the sample is equal to or above about 360 pg/mL; (iv) the level of UCH-L1 alone in the sample is equal to or above about 360 pg/mL; or (v) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is equal to or above about 360 pg/mL; 
 b. not elevated when (i) the level of GFAP alone in the sample is below about 30 pg/mL; (ii) the level of UCH-L1 alone in the sample is below about 360 pg/mL; or (iii) the level of GFAP in the sample is below about 30 pg/mL and level of UCH-L1 in the sample is below about 360 pg/mL; or 
 c. requiring the assays for UCH-L1 and GFAP to be repeated when (i) the level of UCH-L1 alone in the sample cannot be determined or is not reported; (ii) the level of GFAP in the sample is below about 30 pg/mL and the level of UCH-L1 in the sample cannot be determined or is not reported; (iii) the level of GFAP alone in the sample cannot be determined or is not reported; (iv) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is below about 360 pg/mL; or (v) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample cannot be determined or is not reported. 
   
     
     
         55 . The system of  claim 54 , wherein the sample is taken within about 13 hours to within about 48 hours, within about 14 hours to within about 48 hours, within about 15 hours to within about 48 hours, within about 16 hours to within about 48 hours, within about 17 hours to within about 48 hours, within about 18 hours to within about 48 hours, within about 19 hours to within about 48 hours, within about 20 hours to within about 48 hours, within about 21 hours to within about 48 hours, within about 22 hours to within about 48 hours, within about 23 hours to within about 48 hours, within about 24 hours to within about 48 hours, 25 hours to within about 48 hours, within about 26 hours to within about 48 hours, within about 27 hours to within about 48 hours, within about 29 hours to within about 48 hours, within about 30 hours to within about 48 hours, within about 31 hours to within about 48 hours, within about 32 hours to within about 48 hours, within about 33 hours to within about 48 hours, within about 34 hours to within about 48 hours, within about 35 hours to within about 48 hours, within about 36 hours to within about 48 hours, within about 37 hours to within about 48 hours, within about 38 hours to within about 48 hours, within about 39 hours to within about 48 hours, within about 40 hours to within about 48 hours, or within about 12 hour to within about 48 hours after an actual or suspected injury to the head. 
     
     
         56 . (canceled) 
     
     
         57 . The system of  claim 54 , wherein the sample is obtained:
 (a) after the subject sustained an injury to the head caused by physical shaking, blunt impact by an external mechanical or other force that results in a closed or open head trauma, one or more falls, explosions or blasts or other types of blunt force trauma;   (b) after the subject has ingested or been exposed to a chemical, toxin or combination of a chemical and toxin; or   (c) from a subject that suffers from an autoimmune disease, a metabolic disorder, a brain tumor, hypoxia, a viral infection, a fungal infection, a bacterial infection, meningitis, hydrocephalus, or any combinations thereof.   
     
     
         58 . (canceled) 
     
     
         59 . (canceled) 
     
     
         60 . (canceled) 
     
     
         61 . The system of  54 , wherein the assay is (a) an immunoassay or a clinical chemistry assay; or (b) a single molecule detection assay or a point-of-care assay. 
     
     
         62 . (canceled) 
     
     
         63 . The system of  claim 54 , wherein the amount of the at least one sample is: (a) about 10 μL to about 30 μL; or (b) about 20 μL. 
     
     
         64 . (canceled) 
     
     
         65 . The system of  claim 54 , wherein the at least one assay for UCH-L1, at least one assay for GFAP, or at least one assay for UCH-L1 and at least one assay for GFAP is performed in (a) about 10 to about 20 minutes; or (b) about 15 minutes. 
     
     
         66 . (canceled) 
     
     
         67 . (canceled) 
     
     
         68 . The system of  claim 54 , wherein the sample is selected from the group consisting of a whole blood sample, a serum sample, and a plasma sample.